NEURONAL-GLIAL INTERACTIONS IN CNS DEVELOPMENT

拟议的研究计划的长期目标是开发 特异症分子机制研究的模型系统 脑发育过程中神经元-神经胶质细胞的相互作用及其后果 两种细胞类型分化的这些相互作用，包括 对发育调节基因表达的影响。 拟议项目的短期目标和具体目标涉及 利用细胞培养系统研究特定细胞之间的相互作用 由递质表型鉴定的胚胎神经元类型 (5-羟色胺能、多巴胺能、GABA能)和成熟的单层 星形胶质细胞、成纤维细胞或胚胎胶质细胞(放射状胶质细胞、 星形胶质细胞)。这种神经元-神经胶质细胞的相互作用对 这些识别递质的神经元的分化和存活将 用生化方法测量(高亲和力摄取~3H-递质) 和形态(细胞数量定量和形态计量学 免疫细胞化学染色的神经元)。细胞黏附试验，目前 正在开发中，将用于量化这些神经元的黏附 (放射性标记有它们自己的~3H-递质)到相同的非神经元 上述单层。据推测，明显的粘附力 偏好可能产生于特定人群之间的相互作用 同一大脑中识别递质的神经元和未成熟的神经胶质细胞 区域或孕龄。 在未来的实验中，我们希望使用黏附试验来测试可能的 介导这些神经元-胶质细胞相互作用的分子和细胞培养 检测这些相互作用对基因表达的影响的系统 神经元和神经胶质细胞都使用探针来传递细胞特有的信息。

The long-term objectives of the proposed research program are to develop model systems for the study of molecular mechanisms of specific neuronal-glial interactions during brain development, and the consequences of these interactions for the differentiation of both cell types, including effects on the expression of developmentally regulated genes. The short-term goals and specific aims of the proposed project involve the use of a cell culture system to study the interactions between specific types of embryonic neurons, identified by transmitter phenotype (serotonergic, dopaminergic, GABAergic) and monolayers of mature astrocytes, fibroblasts, or embryonic glial cells (radial glia, astrocytes). The effects of such neuronal-glial interactions on the differentiation and survival of these transmitter-identified neurons will be measured both biochemically (high affinity uptake of 3H-transmitters) and morphologically (quantitation of cell number and morphometry of immunocytochemically stained neurons). A cell adhesion assay, currently under development, will be used to quantitate the adhesion of these neurons (radiolabelled with their own 3H-transmitter) to the same non-neuronal monolayers described above. It is hypothesized that distinct adhesion preferences may emerge from interactions between specific populations of transmitter-identified neurons and immature glial cells from the same brain region or gestational age. In future experiments, we hope to use the adhesion assay to test possible molecules mediating these neuronal-glial interactions, and the cell culture system to examine the effects of these interactions on gene expression in both neurons and glia using probes to cell-specific messages.